贵州中医药大学

To investigate the effects of different fermentation conditions on the microbial community structure and chemical composition of Huafengdan Yaomu, this study compared the physicochemical properties, microbial composition, and chemical composition under fermentation with controlled temperature and humidity with those under natural fermentation. The results show the darkening of Yaomu color, significant pH changes, and decreases in the contents of total alkaloids and total flavonoids after fermentation(P<0.05). Microbial community analysis reveals that the number of bacterial amplicon sequence variants(ASVs), abundance, and diversity are significantly increased under fermentation with controlled temperature and humidity, with dominant genera including Ligilactobacillus, Levilactobacillus, and Pichia. In contrast, the natural fermentation group is dominated by Levilactobacillus, Lactiplantibacillus, and Pichia. 584 differential components are screened from 3 003 compounds by metabolomic analysis, including 40 alkaloids and 31 flavonoids. The fermentation group with controlled temperature and humidity exhibits the highest number of differential compositions, with the number of 25 alkaloids including hydropeimine and deltaline significantly increasing, 15 alkaloids including neoline significantly decreasing, 18 flavonoids including loquatoside increasing, and 13 flavonoids including kaempferol decreasing. Natural fermentation is conducive to the production of flavonoid compositions such as kaempferol and quercetin, whereas its effect on reducing toxic alkaloids is less pronounced compared to the fermentation with controlled temperature and humidity. Correlation analysis further indicates that alkaloid compositions are positively correlated with bacterial genera such as Lactiplantibacillus, while flavonoid compositions are positively correlated with fungal genera such as Wickerhamomyces. In conclusion, fermentation conditions significantly affect the chemical composition of Yaomu by regulating the microbial community structure. Fermentation with controlled temperature and humidity demonstrates greater advantages in reducing toxic components, providing a scientific basis for optimizing traditional fermentation processes and improving the quality control of TCM.

To investigate the effect and underlying mechanism of Modified Qinghao Biejia Decoction(MQHBJD) medicated serum in inducing ferroptosis in acute myeloid leukemia(AML) cells, New Zealand rabbits were administered by intragastric gavage to prepare MQHBJD-containing serum and blank serum. After determining the experimental concentrations and incubation time of MQHBJD and the ferroptosis inhibitor Fer-1 by CCK-8 assay, MOLM-13 cells were divided into five groups: blank, 2.5% MQHBJD-containing serum, 5% MQHBJD-containing serum, 10% MQHBJD-containing serum, and 5% MQHBJD-containing serum + Fer-1. Related indicators were detected after 24 hours of intervention. The results showed that, compared with the blank group, the cell inhibition rates in all MQHBJD-containing serum groups were significantly higher(P<0.05, P<0.01, P<0.001), with the 10% MQHBJD-containing serum group exhibiting the highest rate, however, there was no significant difference between this group and the 5% MQHBJD-containing serum group. Levels of malondialdehyde(MDA) and ferrous ions(Fe~(2+)) in the cell culture supernatant were significantly elevated(P<0.05, P<0.01, P<0.001), while glutathione(GSH) levels were significantly decreased(P<0.001); the fluorescence area of reactive oxygen species(ROS) gradually increased(P<0.001), mitochondria exhibited marked atrophy and a reduction in cristae, and ferritin heavy chain 1(FTH1) mRNA and protein expression were significantly decreased(P<0.05, P<0.01, P<0.001), while transferrin receptor 1(TFR1), acyl-CoA synthase long-chain family member 4(ACSL4), and lysosomal phosphatidylcholine acyltransferase 3(LPCAT3) mRNA and protein expression significantly increased(P<0.05, P<0.01, P<0.001). Compared with the 5% MQHBJD-containing serum group, the cell inhibition rate was significantly lower in the 5% MQHBJD-containing serum + Fer-1 group(P<0.001), MDA and Fe~(2+) levels were significantly reduced(P<0.05, P<0.001), GSH levels were significantly increased(P<0.01), the ROS fluorescence area was significantly decreased(P<0.001), mitochondrial atrophy and reduced cristae were improved, FTH1 protein expression was significantly increased(P<0.01), and TFR1, ACSL4, LPCAT3 protein expression were significantly decreased(P<0.001). In conclusion, MQHBJD can effectively induce ferroptosis in AML cells, and its mechanism may be related to the regulation of the TFR1/ACSL4/LPCAT3 signaling axis.

Objective To investigate the effect of electroacupuncture(EA)on low-grade duodenal inflammation and gastrointestinal motility in rats with functional dyspepsia(FD)and the underlying mechanism involving the Gas6/MerTK efferocytosis pathway.Methods Male SD rats were randomly divided into control,model,EA+Gas6 overexpression,EA+Gas6 inhibitor,and EA groups.The EA+Gas6 overexpression group received adenovirus-mediated Gas6 overexpression,the EA+Gas6 inhibitor group received a Gas6 inhibi-tor,and the EA group received EA treatment.Pro-inflammatory and anti-inflammatory cytokine levels in duodenal tissue were quanti-fied using ELISA.Protein expression of Gas6,MerTK,and efferocytosis pathway-related indicators(SIRPα,Axl,CD47,MFGE8,Pros)was assessed by Western blot(WB).Results Compared with the control group,the model group showed significantly increased expression of IL-17 and IFN-γ(P<0.001)and decreased expression of IL-4 and TGF-β(P<0.001)in the duodenum.The expression of Gas6 and MerTK was reduced(P<0.001),efferocytosis was impaired,and protein expression of efferocytosis pathway-related indica-tors was decreased(P<0.001).Compared with the model group,both the EA+Gas6 overexpression group and the EA group exhibited decreased IL-17 and IFN-γ expression(P<0.05),increased IL-4 and TGF-β expression(P<0.05),enhanced efferocytosis,and increased protein expression of Gas6,MerTK,and efferocytosis pathway-related indicators(P<0.05).Compared with the EA group,the EA+Gas6 overexpression group showed further decreases in IL-17 and IFN-γ expression(P<0.05),increases in IL-4 and TGF-β expression(P<0.05),slight enhancement of efferocytosis(P<0.05),and elevated protein expression of Gas6,MerTK,and efferocyto-sis pathway-related indicators(P<0.05).In contrast,the EA+Gas6 inhibitor group demonstrated opposite trends.Conclusion EA may alleviate low-grade duodenal inflammation,enhance efferocytosis,and promote gastrointestinal motility in FD rats,possibly through the Gas6/MerTK pathway.

Codonopsis javanica, a Campanulaceae herb endemic to Guizhou, is locally called "Tudangshen", its root is used in medicine to fortify spleen-qi, moisten the lung and arrest cough, and promote lactation. In June 2023, root rot of C. javanica was observed in a cultivation field in Xiuwen County (106°46'48" E, 26°53'59" N), Guizhou Province, with the estimated disease incidence ranging from 20% to 50%. The initial symptoms were light brown spots on the surface of the root, and the spots were brown or black in severe cases, and the lower end of the root and the rhizome were rotted. The aboveground portions of the leaves exhibited yellowing and wilting, followed by withering and death. The diseased root tissue (5 mm × 5 mm) was cut and disinfected with 75 % alcohol for 15 s, 5 % sodium hypochlorite treatment for 1 min, followed by three rinses with sterile distilled water. After drying, the samples were transferred onto Potato Dextrose Agar (PDA) and incubated at 28 °C in darkness for 3-5 days. Through hyphal tip culture methodology (Senanayake et al. 2020), 12 strains with similar morphology were obtained. The colonies were white to purplish red. The fungus produced two types of conidia: microconidia and macroconidia. The microconidia were ovate, without septa or 1-2 septa, and the size was 4.98-9.22 × 1.88-3.65 μm. The macroconidia were sickle-shaped, slightly curved, 1-4 septa, and the size was 21.55-37.92 × 3.10-6.50 μm. Chlamydospores were subspherical to spherical. Based on these morphological features, the isolates were suspected to be Fusarium oxysporum (Leslie et al. 2006). For molecular confirmation of the morphological identification, strain ZPJ1 was randomly selected as a representative isolate for DNA extraction. The primers ITS1/ITS4, EF1-F6/R6, and RPB2-5F/RPB2-7cR were used to amplify three genomic DNA regions including the internal transcribed spacer (ITS)(Crous et al. 2021), translation elongation factor 1-alpha (EF1)(Moslemi et al. 2017), and RNA polymerase II second largest subunit (RPB2)(Xiang et al. 2025). The sequences of ZPJ1 (accession numbers PV750775, PV759349, and PV759350 for ITS, EF1 and RPB2, respectively) were deposited in GenBank, demonstrating a sequence homology of 99 % to 100 % (546/546, 702/705, 974/978) with F. oxysporum (accession numbers PP596264, MN892354, MN457466). Based on morphological features and maximum likelihood method analyses of combined ITS, EF1, and RPB2 gene sequences, strain ZPJ1 was identified as F. oxysporum. Pathogenicity experiments were conducted in a greenhouse with a temperature ranging from 24 to 27 °C and a relative humidity of 80 %. Root wounding was performed on potted C. javanica seedlings (n = 5, 1-year-old) using a sterile needle. A spore suspension (150 mL, 1 × 106 conidia/mL) of strain ZPJ1 was inoculated onto the wounds, with an equal volume of sterile water used as the control. After 18 days, all inoculated plants developed characteristic root rot symptoms, while the plants in the control group remained healthy and did not show any symptoms of root rot disease. F. oxysporum was successfully reisolated from the symptomatic roots and identified by the morphology and the molecular methods de-scribed above, meeting Koch’s postulates. Based on current understanding, this is the first report of F. oxysporum causing root rot in C. javanica in China. These findings provide a basis for the early detection of C. javanica root rot caused by F. oxysporum and the formulation of targeted control measures.